Identification, molecular characterization, and structural analysis of the blaNDM-1 gene/enzyme from NDM-1-producing Klebsiella pneumoniae isolates

Melgarejo, Jhessyca Leal; Cardoso, Marlon Henrique; Pinto, Ingrid Batista; Faria-Junior, Célio; Mendo, Sónia; Oliveira, Carina Elisei de; Franco, Octávio Luiz

doi:10.1038/s41429-018-0126-z

Cited by 6 publications

(7 citation statements)

References 41 publications

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“…NDM-producing bacteria are resistant to nearly all accessible antibiotics except polymyxin E, but mcr-1 emerging as an element of resistance limits the number of available therapeutic options to this last resort antibiotic [11]. The bla NDM-1 -resistance gene encodes the NDM-1 enzyme and is traced in mobile genetic structures, mainly plasmids, which can facilitate bla NDM-1 dissemination [12]. bla NDM-1 plasmids are isolated from various bacterial species, demonstrating the significant role of plasmids in spreading bla NDM-1 [10].…”

Section: Introductionmentioning

confidence: 99%

Detection of NDM-1 producing Klebsiella pneumoniae ST15 and ST147 in Iran during 2019–2020

Rad

Goudarzi

et al. 2021

AMicr

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Carbapenems are employed to treat infections caused by Gram-negative bacteria including Klebsiella pneumoniae. This research is aimed to perform phenotypic detection of β-lactamases and molecular characterization of NDM-1 positive K. pneumoniae isolates. Another objective is to investigate NDM-1 producing K. pneumoniae among children in Iran. From 2019 to 2020, altogether 60 K. pneumoniae isolates were acquired from various patients in certain Iranian hospitals. Antimicrobial susceptibility testing was performed by disk diffusion and broth microdilution methods. In addition, mCIM and eCIM were used to confirm the production of carbapenemases and metallo-beta-lactamases (MBLs), respectively. Detection of resistance genes namely, blaNDM-1, blaIMP, blaVIM, blaKPC, blaOXA-48-like, blaCTX-M, blaSHV, blaTEM, and mcr-1 was performed by PCR and confirmed by DNA sequencing. Multilocus sequence typing (MLST) was employed to determine the molecular typing of the strains. According to the findings, the highest rate of carbapenem resistance was detected against doripenem 83.3% (50). Moreover, 31.7% (19) were resistant to colistin. Further to the above, altogether 80% (48) were carbapenemase-producing isolates and among them 46.7% (28) of the isolates were MBL and 33.3% (20) isolates were serine β-lactamase producer. According to the PCR results, 14 isolates produced blaNDM-1. Remarkably, four blaNDM-1 positive isolates were detected in children. In addition, these isolates were clonally related as determined by MLST (ST147, ST15). Altogether ten blaNDM-1 positive isolates were ST147 and four blaNDM-1 positive isolates were ST15. Based on the results, the emergence of NDM-producing K. pneumoniae among children is worrying and hence, it is necessary to develop a comprehensive program to control antibiotic resistance in the country.

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Section: Introductionmentioning

confidence: 99%

Detection of NDM-1 producing Klebsiella pneumoniae ST15 and ST147 in Iran during 2019–2020

Rad

Goudarzi

et al. 2021

AMicr

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“…21 Most Enterobacteriaceae are healthy flora; however, some become pathogens, causing a diversified severe infection, including systemic bacteremia, communityacquired infections, healthcare-associated infections (HAIs), and both complicated and uncomplicated urinary tract infections (UTIs). 22,23 Klebsiella pneumoniae remains the leading pathogen producing carbapenemase 24 and MBL enzymes 17,25 and has been reported to show co-resistance through containing multiple ESBL genes simultaneously. 26 Some antibiotic resistance genes (ARGs) accumulate multiple random mutations in their gene sequences over time and become dysfunctional resistance genes (resistance pseudogenes) that fail to exhibit expected resistance phenotypes.…”

Section: Introductionmentioning

confidence: 99%

Genotypic to Phenotypic Resistance Discrepancies Identified Involving β-Lactamase Genes, blaKPC, blaIMP, blaNDM-1, and blaVIM in Uropathogenic Klebsiella pneumoniae

Urmi¹,

Nahar²,

Rana³

et al. 2020

IDR

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Introduction: Klebsiella pneumoniae carbapenemase (KPC) belongs to the Group-A βlactamases that incorporate serine at their active site and hydrolyze various penicillins, cephalosporins, and carbapenems. Metallo-beta-lactamases (MBLs) are group-B enzymes that contain one or two essential zinc ions in the active sites and hydrolyze almost all clinically available β-lactam antibiotics. Klebsiella pneumoniae remains the pathogen with the most antimicrobial resistance to KPC and MBLs. Methods: This research investigated the blaKPC, and MBL genes, namely, blaIMP, blaVIM, and blaNDM-1 and their phenotypic resistance to K. pneumoniae isolated from urinary tract infections (UTI) in Bangladesh. Isolated UTI K. pneumoniae were identified by API-20E and 16s rDNA gene analysis. Their phenotypic antimicrobial resistance was examined by the Kirby-Bauer disc diffusion method, followed by minimal inhibitory concentration (MIC) determination. blaKPC, blaIMP, blaNDM-1, and blaVIM genes were evaluated by polymerase chain reactions (PCR) and confirmed by sequencing. Results: Fifty-eight K. pneumoniae were identified from 142 acute UTI cases. Their phenotypic resistance to amoxycillin-clavulanic acid, cephalexin, cefuroxime, ceftriaxone, and imipenem were 98.3%, 100%, 96.5%, 91.4%, 75.1%, respectively. Over half (31/58) of the isolates contained either blaKPC or one of the MBL genes. Individual prevalence of blaKPC, blaIMP, blaNDM-1, and blaVIM were 15.5% (9), 10.3% (6), 22.4% (13), and 19% (11), respectively. Of these, eight isolates (25.8%, 8/31) were found to have two genes in four different combinations. The coexistence of the ESBL genes generated more resistance than each one individually. Some isolates appeared phenotypically susceptible to imipenem in the presence of blaKPC, blaIMP, blaVIM, and blaNDM-1 genes, singly or in combination. Conclusion: The discrepancy of genotype and phenotype resistance has significant consequences for clinical bacteriology, precision in diagnosis, the prudent selection of antimicrobials, and rational prescribing. Heterogeneous phenotypes of antimicrobial susceptibility testing should be taken seriously to avoid inappropriate diagnostic and therapeutic decisions.

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Section: Introductionmentioning

confidence: 99%

“…The excessive and indiscriminate use of beta-lactam antimicrobials has culminated in the emergence of antibiotic-resistant K. pneumoniae strains and other carbapenem-resistant Enterobacteria 1 , 5 , 6 , 7 . The development of resistance is related to the production of beta-lactamases which is mediated by conjugative plasmids 8 , 9 , 10 , 11 , 12 .…”

Section: Introductionmentioning

confidence: 99%

“…Additionally, resistance to carbapenems can occur due to the production of other enzymes, such as metallo-beta-lactamases, e.g., the New Delhi metallo-beta-lactamase-1 (NDM-1). Since its detection in 2008 in New Delhi, India 23 strains producing NDM-1 have been reported in many countries, including Brazil 6 , 8 , 9 , 24 , 25 . Considering the importance of knowing which resistance genes and plasmids are circulating among multi-drug resistant clinical isolates in hospitals in Brazil, we investigated the bla KPC , bla GES , bla NDM , bla VIM , and bla IMP genes, and the most common plasmid Incs in K. pneumoniae (FIB, Q, A/C, L/M, N, HI2, and HI1B) to analyze the clonal relationship between KPC resistant clinical isolates obtained from a public hospital in Recife-PE, Brazil.…”

Section: Introductionmentioning

confidence: 99%

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High plasmid variability, and the presence of IncFIB, IncQ, IncA/C, IncHI1B, and IncL/M in clinical isolates of Klebsiella pneumoniae with bla KPC and bla NDM from patients at a public hospital in Brazil.

Oliveira

Beltrão

Scavuzzi

et al. 2020

Rev. Soc. Bras. Med. Trop.

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Introduction: Antibiotic resistance in carbapenemase-producing Klebsiella pneumoniae is acquired and disseminated mainly by plasmids. Therefore, we aimed to investigate the occurrence of carbapenemase genes, analyze the genetic diversity by ERIC-PCR, and examine the most common plasmid incompatibility groups (Incs) in clinical isolates of K. pneumoniae from colonization and infection in patients from a hospital in Brazil. Methods: Twenty-seven isolates of carbapenem-resistant K. pneumoniae were selected and screened for the presence of carbapenemase genes and Incs by PCR, followed by amplicon sequencing. Results: The bla KPC and bla NDM genes were detected in 24 (88.8 %) and 16 (59.2 %) of the isolates, respectively. Thirteen isolates (48.1 %) were positive for both genes. The IncFIB (92.6 %) and IncQ (88.8 %) were the most frequent plasmids, followed by IncA/C, IncHI1B, and IncL/M, indicating that plasmid variability existed in these isolates. To our knowledge, this is the first report of IncHI1B in Brazil. We found eight isolates with clonal relationship distributed in different sectors of the hospital. Conclusions: The accumulation of resistance determinants, the variability of plasmid Incs, and the clonal dissemination detected in K. pneumoniae isolates demonstrate their potential for infection, colonization, and the dissemination of different resistance genes and plasmids.

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Identification, molecular characterization, and structural analysis of the blaNDM-1 gene/enzyme from NDM-1-producing Klebsiella pneumoniae isolates

Cited by 6 publications

References 41 publications

Detection of NDM-1 producing Klebsiella pneumoniae ST15 and ST147 in Iran during 2019–2020

Detection of NDM-1 producing Klebsiella pneumoniae ST15 and ST147 in Iran during 2019–2020

<p>Genotypic to Phenotypic Resistance Discrepancies Identified Involving β-Lactamase Genes, <em>bla</em>KPC, <em>bla</em>IMP, <em>bla</em>NDM-1, and <em>bla</em>VIM in Uropathogenic <em>Klebsiella pneumoniae</em></p>

High plasmid variability, and the presence of IncFIB, IncQ, IncA/C, IncHI1B, and IncL/M in clinical isolates of Klebsiella pneumoniae with bla KPC and bla NDM from patients at a public hospital in Brazil.

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